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. 2019 Mar 6;9:3633. doi: 10.1038/s41598-019-40002-0

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Identification of the signaling pathway responsible for lithocholic acid (LCA)-induced effects in H9c2 cells incubated in high-glucose medium. (A) Effects of LCA on the SERCA2 mRNA levels modified by high glucose in H9c2 cells were blocked by the protein kinase A (PKA) inhibitors PKAI and H-89, but not by the Epac inhibitor ESI-09 (n = 6). (B) Representative image of Western blots indicating the changes in protein levels of TGR5, SERCA2, phospholamban (PLN), and phosphorylated PLN (PPLN) using Actin as an internal control (n = 4). (C) The upper panel shows the variations in TGR5/Actin expression between the 6 groups, the middle panel compares the differences in PPLN/PLN expression between the 6 groups, and the lower panel shows changes in SERCA2/Actin expression between the 6 groups. The results indicate that cells treated with high glucose that improved with LCA treatment were blocked by the PKA inhibitors but not by the Epac inhibitor (n = 4), (* p-value < 0.05, **p-value < 0.01 versus control group; ^#p-value < 0.05, ^##p-value < 0.01 versus vehicle group).